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为了实现增强型绿色荧光蛋白基因(egfp)在生防真菌淡紫拟青霉9410菌株中的转化,借助中间质粒pcDNA3.1(-)构建nptⅡ-egfp融合基因的表达载体pUPNGT,然后采用根癌农杆菌介导的转化法将egfp基因转化到淡紫拟青霉9410菌株中。PCR检测和Southern blotting分析结果表明,egfp基因以单拷贝形式整合到淡紫拟青霉9410的基因组中。荧光显微镜观察结果显示,转化子在488nm光源的激发下能产生绿色荧光。这些结果说明egfp基因已成功转化至淡紫拟青霉9410菌株并获得表达。这些工作可为淡紫拟青霉在不同条件下的防效评价、环境安全评价等提供新的途径和方法。
In order to achieve the transformation of egfp gene into the antifungal Paecilomyces lilacin 9410 strain, the expression vector pUPNGT of nptⅡ-egfp fusion gene was constructed by using the intermediate plasmid pcDNA3.1 (-). The egfp gene was transformed into Paecilomyces lilacinus 9410 by Agrobacterium-mediated transformation. PCR analysis and Southern blotting analysis showed that the egfp gene was integrated into the genome of Paecilomyces lilacinus 9410 in a single copy. Fluorescence microscopy showed that transformants were able to generate green fluorescence when excited by a 488 nm light source. These results indicate that the egfp gene has been successfully transformed into Paecilomyces lilacinus 9410 strain and expressed. These works can provide new ways and methods for Paecilomyces lilacinus in different conditions of the effectiveness evaluation, environmental safety assessment.